Targeted lipidomics on HT-29 cells that were treated with and without propionate

Targeted lipidomics was performed on HT-29 cells that were treated with and without propionate. The goal was to assess how lipids were impacted with priopionate treatment. The experimental design included treating HT-29 cells (approximately 214800/well) with and without 0.2 mM propionate for 48 hours. The treatment media was EBSS, 5% FBS and p/s. There were four biological replicates for the control and treatment cohorts. Total lipid extracts from cell pellets were prepared using a modified MTBE lipid extraction protocol. Total lipid extracts were analyzed by liquid chromatography coupled to targeted tandem mass spectrometry (LC-MS/MS). The LC-MS/MS analyses were performed on an Ultimate 3000 Ultra High-Performance Liquid Chromatograph coupled to a Thermo TSQ Altis Tandem Quadrupole Mass Spectrometer (Thermo Scientific, San Jose, CA). LC-MS/MS methodology was adapted from the literature. LC-MS/MS data was acquired using Thermo’s Xcalibur software and data processing was achieved using Xcalibur 4.2 and TraceFinder 5.1.

本研究针对经丙酸酯处理与未处理的HT-29细胞开展靶向脂质组学（targeted lipidomics）分析，旨在评估丙酸酯处理对细胞脂质谱的影响。实验方案为：将每孔接种约214800个的HT-29细胞，分别用含0.2 mM丙酸酯与不含丙酸酯的培养基处理48小时；处理培养基为添加5%胎牛血清（FBS）与双抗（p/s，即青霉素-链霉素）的Earle平衡盐溶液（EBSS），对照组与处理组各设置4次生物学重复。本研究采用改良的甲基叔丁基醚（MTBE）脂质提取方案对细胞沉淀中的总脂质进行提取，随后通过液相色谱联用靶向串联质谱（LC-MS/MS）开展分析：LC-MS/MS分析在联用赛默飞TSQ Altis三重四极杆质谱仪（Thermo Scientific，美国加州圣何塞）的Ultimate 3000超高效液相色谱仪上完成，分析方法改编自已发表文献。LC-MS/MS数据采集采用赛默飞Xcalibur软件，数据处理则通过Xcalibur 4.2与TraceFinder 5.1软件实现。